Concerning the ONCO-ND1 line, the use of immunological checkpoint inhibitors has highlighted the great potential of these therapies immuno-oncological therapies characterised by positive and lasting responses never observed before. In many cases, these types of treatments allow both complete remission of the disease and increased survival and sometimes cure of the neoplasm even at an advanced stage. However, one of the major problems is that only a percentage of patients (typically 25% in all types of immune-responsive neoplasms) are sensitive to such treatment strategies. It is therefore imperative to identify the patients who can benefit most from these types of therapies, which are often expensive and burdened with side effects. Furthermore, there is an urgent call for the development of innovative immunotherapeutic strategies capable of optimising the therapeutic effects and benefits in human malignancies. At present, despite correlations between gene expression parameters, no single molecular parameters of the malignancy are known that can accurately predict response to immunotherapy, and the diagnostic parameters used to date generally have a low sensitivity and specificity that often contribute to the failure of response prediction. This results in a loss of a proportion of patients who might respond to treatment and exposure to side effects that do not
translatable into therapeutic benefit in another share of patients. Another major limitation is the need to predict the side effects of this type of treatment, which can sometimes even be fatal. To this end, we recently highlighted how the expression of certain HLA haplotypes are predictive of immune checkpoint inhibitor-induced pneumonias (Correale et al Cells 2020). The identification of tumour and host genetic patterns
in the prediction of toxicity to anti-cancer therapies is another major knot to be unravelled in the optimisation of anti-cancer therapeutic strategies.
The molecular/computational platform ONCO-ND1 developed by IMMUNOMICA, which integrates the research results of the proposing group as evidenced by numerous international discoveries and publications, intends to specifically address these two objectives.

The ONCO-ND1 platform enables the overall analysis of the immunological characteristics of the tumour and its microenvironment in each individual neoplastic specimen. The methodologies developed by the group of proponents have the characteristic of maximising the data generated from each individual sample and on the other of optimising its interpretation by adopting a single acquisition methodology based on Next Generation Sequencing to obtain an accurate immunological characterisation that would traditionally require different assays as well as different formats and modes of interpretation.
By means of a single sequencing acquisition of the DNA/RNA sample under investigation, applying the algorithms developed by the proposing researchers, it will be possible to identify:

• A Tumour Mutational Burden (associated with response to immunotherapy)
• B Neoantigen load (associated with response to immunotherapy)
• C Presence of Microsatellite Instability (MSI) (strong predictor of response to immunotherapy)
• D T-cell receptor repertoire (high clonality is associated with response to immunotherapy)
• E HLA subtype (presence of certain HLA haplotypes is associated with high response to immunotherapy)
• F Immune signature expressions of immune response and resistance (associated with response or resistance to immunotherapy)
• G The presence of specific somatic tumour mutations associated with immunotherapy resistance (specific mutations in the interferon pathway and other mutations such as those inherent in the EGFR/MAPK pathways are associated with resistance to immunotherapy)
• H The presence of germline polymorphisms associated with response
• I The composition of the microbiome (presence of specific bacterial strains are associated with response to immunotherapy)
• L Expression from peripheral blood of miRNA signatures predictive of response to immunotherapy

The innovation of the ONCO-ND1 platform is twofold, on the one hand the innovative methods of DNA/RNA extraction from tumour tissue and biological fluids and characterisation of the tumour microenvironment from paraffin-embedded samples, and on the other hand the algorithms for analysing sequencing data developed over the years by a collaboration between Prof. Bedognetti’s group at SIDRA and the University of Turin.
Bedognetti at the SIDRA Medical Research Centre in Doha (Qatar) and Prof. Ceccarelli at the BIOGEM Research Centre. In particular, for each parameter analysed by the platform, specific scientific publications have demonstrated their association with the response to immunotherapy. While the
most research was evaluated one single parameter at a time, the proposed platform allows the evaluation of all parameters organised in 10 categories with a single integrated DNA and RNA analysis test. The predictive synergism of these parameters, e.g. of immune signatures + mutational burden, or germline polymorphisms such as HLA and Fc Receptors and tumour mutational burden has recently been demonstrated. The guidelines
of the leading cancer immunotherapy society (The Society For The Immunotherapy of Cancer) (Bedognetti, Ceccarelli, et al Journal for Immunotherapy of Cancer. 2019 May 22;7(1):131.) emphasised the importance of integrated assessment of these parameters for future implementation of personalised immunotherapy.
The technologies developed for immunological characterisation of tumours from sequencing data feeding into the ONCO-ND1 platform could be used for the definition of personalised therapies and for the selection of patients most suitable for immunotherapies. The competences developed by Prof. Michele Caraglia’s team in Molecular Oncology and Precision Medicine will be instrumental in defining the diagnostic reports and the associations of immunological/molecular characteristics with the most appropriate therapeutic strategies and clinical trials for each individual sample. In the same vein, the integrated use of multiple genetic and molecular markers could lead to the clear definition of the risk of developing severe side effects (such as fatal interstitial pneumonitis) while avoiding the associated risk of occurrence.
Ultimately, IMMUNOMICA’s goal is to fill a gap in the market, especially in Italy and Europe, where, although there are now a large number of centres offering sequencing services, no specialised and analytical market solutions focused on immunotherapies have been implemented. The international relevance of the skills and technologies developed by the group of proposers places IMMUNOMICA at the frontier of the scientific and technological state of the art in this field. A further success factor of this initiative is that it is a spin-off of the BIOGEM Research Centre in Ariano Irpino, where IMMUNOMICA will be located, with the possibility of access to molecular biology, sequencing, high-performance computing and animal model laboratories among the best equipped in Italy. It should also be emphasised that the research teams of Prof. Michele Ceccarelli and Prof. Michele Caraglia are already working at the Biogem research centrer.